Pharmaceutical composition for treating cancer in patient having specific gene marker

Abstract

The present invention provides a pharmaceutical composition for use in the treatment of a cancer disease in a subject, comprising O-(5-amino-2-phenylbenzoxazol-7-yl) methyl-3, 5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof, the pharmaceutical composition being administered to the subject having a non-fast (slow and / or medium speed) type NAT2 gene.

Description

[0001] This application is a divisional application of a patent application with an application date of August 31, 2020, application number 202080062102.X, and invention name “Pharmaceutical composition for cancer treatment of patients with specific genetic markers”. Technical Field

[0002] The present invention relates to a pharmaceutical composition comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof which is most suitable for patients with cancer or the like having specific gene markers.

[0003] Furthermore, the present invention relates to a method for treating cancer, etc., comprising the step of administering a pharmaceutical composition containing O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof to a patient with cancer, etc. having a specific gene marker. Background Art

[0004] Since individual differences in isoniazid neurotoxicity are caused by genetic differences in N-acetylation ability, N-acetylation polymorphisms were discovered more than 50 years ago. In addition to isoniazid, many aromatic amine drugs such as sulfamethazine are affected by acetylation polymorphisms, affecting the therapeutic effects and toxicity of many therapeutic agents. N-acetyltransferase 2 (NAT2) is an enzyme that transfers the acetyl group of acetyl-CoA to aromatic amine-containing compounds, which are substrates of the enzyme. NAT2 is encoded by an 870-bp gene (NAT2), and genetic polymorphisms of NAT2 are common in humans.

[0005] Furthermore, it is known that there are three types of NAT2 acetylation speed: rapid, intermediate, and slow (SNP: single nucleotide polymorphism) (Pharmacogenomics, Vol. 13: pp. 31-41, 2012). These types can be distinguished by their gene polymorphisms.

[0006] Genetic polymorphisms of NAT2 are known to alter the efficacy and toxicity of many arylamine and hydrazine drugs and increase the risk of many arylamine carcinogen-related cancers. In addition, drugs such as sulfasalazine (SSZ) are known to be acetylated by NAT2, but side effects are often more likely to occur in patients with slow acetylation (J Rheumatol. 2002, Dec; 29(12):2492-2499) (Genotyping the NAT2 gene followed by estimation of diplotype configuration before administration of SSZ is likely to reduce the frequency of adverse effects in Japanese patients with RA. http: / / www.jrheum.org / content / 29 / 12 / 2492).

[0007] There are seven representative NAT2 gene polymorphisms: (i) 191G>A (rs1801279), (ii) 282C>T (rs1041983), (iii) 341T>C (rs1801280), (iv) 481C>T (rs1799929), (v) 590G>A (rs1799930), (vi) 803A>G (rs1208), and (vii) 857G>A (rs1799931). Among them, SNPs (i), (iii), (v), (vi), and (vii) resulted in amino acid changes (i.e., (i) 191G>A from arginine to glutamate, (iii) 341T>C from isoleucine to threonine, (v) 590G>A from arginine to glutamine, (vi) 803A>G from lysine to arginine, and (vii) 857G>A from glycine to glutamate).

[0008] On the other hand, biliary tract cancer has been increasing in Japan in recent years. The number of deaths in 2016 was approximately 18,000, making it the seventh leading cause of cancer death. Biliary tract cancer has different treatment strategies and chemotherapy effects depending on the site of occurrence. As chemotherapy, the combination of gemcitabine and cisplatin is currently used as standard chemotherapy. However, there is currently no established second-line treatment method recommended for patients with biliary tract cancer who are ineffective with standard chemotherapy, and there is a high clinical need for chemotherapy as a second-line treatment method.

[0009] O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine (hereinafter, sometimes referred to as "JPH203") has a selective inhibitory effect on L-amino acid transporter 1 (LAT1) specifically expressed in cancer cells, and is therefore expected to be effective as a new antitumor agent (Patent Document 1). Currently, JPH203 is being tested in a Phase II trial in Japan on patients with biliary tract cancer by the applicant.

[0010] Prior art literature Patent Literature Patent Document 1: International Publication No. 2008 / 081537 Non-patent literature Non-patent literature 1: Pharmacogenomics, Vol. 13: pp. 31-41, 2012 Non-patent literature 2: Drug Metab Pharmacokinet, 2012; 27(1): 155-161 Non-patent document 3: J. Rheumatol, 2002, Dec; 29(12): pp. 2492-2499 Non-patent document 4: Jutabha et al. Journal of Pharmacological Sciences, Volume 130, Issue 3, Supplement (Proceedings of the 89th Annual Meeting of the Japanese Pharmacological Society) 3-O-84 Summary of the invention

[0011] Technical issues While studying a new second-line chemotherapy for patients with biliary tract cancer who have failed to respond to standard chemotherapy, the inventors found, based on the results of pharmacokinetic parameter analysis of JPH203 obtained in a Phase I trial in Japan, that among subjects given the same dose per unit body surface area, the individual differences in the concentration of N-acetylated forms in the blood and urine were greater than the concentration of JPH203 in the blood. Further research was continued, and it was found that there was a correlation between the difference in acetylation rate in the genetic polymorphism of NAT2 and the safety and efficacy of JPH203, thereby completing the present invention.

[0012] Technical Solution That is, the present invention provides a pharmaceutical composition for use in the treatment of a disease in a subject, comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition is administered to the subject having a non-fast (slow and / or intermediate) type NAT2 gene.

[0013] In addition, the present invention provides a method for treating a disease in a subject, comprising administering to the subject a pharmaceutical composition comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof after the subject is identified as having a non-fast (slow and / or intermediate) type NAT2 gene.

[0014] In addition, the present invention provides a use of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease of a subject, characterized in that after the subject is identified as having a non-fast (slow and / or intermediate) type NAT2 gene, a pharmaceutical composition comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0015] Furthermore, the present invention provides a method for predicting whether O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine exerts an effect on a specific cancer or the like, by determining whether the object has a non-fast (slow and / or medium) type NAT2 gene in the NAT2 gene.

[0016] In addition, the present invention provides the use of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof in the manufacture of a drug for treating a specific disease, which includes determining whether the NAT2 gene of the subject has a non-fast (slow and / or medium) type NAT2 gene before using the drug to treat the specific disease, and predicting whether O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine has an effect on the subject of the specific disease.

[0017] The present invention provides a gene diagnosis kit for determining whether a non-rapid NAT2 gene is present in a subject's NAT2 gene, so as to diagnose whether O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine exerts an effect on a specific disease, and provides a pharmaceutical composition containing O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof, which is used in combination with the kit or NAT2 gene diagnosis.

[0018] The terms "subject" and "patient" are used interchangeably herein, and are preferably human.

[0019] Effects of the Invention The pharmaceutical composition of the present invention can provide treatment for cancer and the like with improved safety of the drug and improved effectiveness of the effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The figure shows the Nac-JPH203 / JPH203 ratio in the blood of rats treated with JPH203 and a NAT2 inhibitor in combination.

[0021] Figure 2 The graph shows the concentration of BPA enclosed in pancreatic cancer-derived T3M4 cells after adding boronated phenylalanine (BPA) and then adding 10 μM JPH203 15 minutes later. DETAILED DESCRIPTION

[0022] Hereinafter, the present invention will be described in detail.

[0023] O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine (JPH203) O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine, which is the active ingredient of the present invention, is a compound disclosed in Patent Document 1, and the applicants are currently conducting a Phase II trial.

[0024] It should be noted that after intravenous administration of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine (JPH203), it is mainly metabolized (acetylated) by liver NAT2 to become the acetyl form (Nac-JPH203), and its physiological activity is greatly reduced (Drug Metab Pharmacokinet. 2012; 27(1): 155-161).

[0025] JPH203 can also be used in the form of its pharmacologically acceptable salt. For example, the amino group in the molecule of JPH203 forms a salt with an acid. Such a salt is not particularly limited, and examples thereof include salts with inorganic acids such as hydrochloric acid and sulfuric acid, and carboxylic acids. Among them, hydrochloride is preferred.

[0026] Pharmaceutical composition The pharmaceutical composition of the present invention comprises JPH203 or a pharmacologically acceptable salt thereof as an active ingredient. In addition, pharmaceutical additives may be included as needed. The pharmaceutical composition of the present invention may be orally administered in the form of solid preparations such as tablets, granules, fine granules, powders, capsules, or liquids, jellies, syrups, or the pharmaceutical composition may be administered parenterally in the form of injections, nasal preparations, suppositories, inhalants, transdermal preparations, etc.

[0027] In particular, since the pharmaceutical composition of the present invention can be used as an anticancer agent, etc., it is preferably formulated as an injection. Examples of such an injection include an injection containing a pH adjuster and cyclodextrins as the active ingredient of the present invention.

[0028] Specific examples of the injection include intravenous, subcutaneous, intradermal, intramuscular injections, and intravenous drip injections.

[0029] Examples of the pH adjuster that can be incorporated into the injection of the present invention include alkali metal hydroxides such as sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide, alkali metal hydrides such as sodium hydride and potassium hydride, and carbonates of alkali metals or alkaline earth metals. Sodium hydroxide and sodium carbonate are particularly preferred, and sodium hydroxide is more preferred.

[0030] The injection can be appropriately adjusted to an appropriate pH using a pH adjuster. The pH of the injection of this embodiment is preferably 3 to 6, more preferably 3 to 5, further preferably 3 to 4.5, and particularly preferably 3.5 to 4.5.

[0031] As cyclodextrins that can be formulated in the injection of the present invention, for example, unmodified cyclodextrin and modified cyclodextrin can be cited. As unmodified cyclodextrin, for example, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, etc. can be cited. In addition, as modified cyclodextrins, for example, dimethyl-α-cyclodextrin, dimethyl-β-cyclodextrin, dimethyl-γ-cyclodextrin, hydroxypropyl-α-cyclodextrin, hydroxypropyl-γ-cyclodextrin, sulfobutyl ether-α-cyclodextrin, sulfobutyl ether-β-cyclodextrin, sulfobutyl ether-γ-cyclodextrin, maltosyl-α-cyclodextrin, etc. can be cited.

[0032] The cyclodextrins may be used alone or in any combination of two or more. From the viewpoint of being able to reduce the number of insoluble particles formed even when dissolved in a non-strongly acidic aqueous solution and improving the resolubility of the lyophilized preparation in a non-strongly acidic aqueous solution, the cyclodextrins are preferably hydroxypropyl-β-cyclodextrin or sulfobutyl ether-β-cyclodextrin, and more preferably sulfobutyl ether-β-cyclodextrin.

[0033] Here, sulfobutyl ether-cyclodextrin is a structure shown in the following formula 1, and the inner side of the ring structure has high hydrophobicity. Therefore, it forms a complex with O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine, which also has high hydrophobicity, through hydrophobic interaction. The "sulfobutyl ether-cyclodextrin complex" mentioned in this specification refers to a complex formed by the hydrophobic interaction as described above.

[0034] [Chemical formula 1]

[0035] Alternatively, O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine as an active ingredient may be used in the form of a sulfobutyl ether-cyclodextrin complex (hereinafter referred to as "JPH203-SBECD") for injection.

[0036] If necessary, a buffer, a suspending agent, a solubilizing agent, a stabilizer, an isotonicity adjusting agent, a preservative, etc. may be further added to the injection of the present invention.

[0037] Examples of the buffer include boric acid buffer, phosphate buffer, citric acid buffer, acetic acid buffer, and Tris buffer.

[0038] Examples of the suspending agent include methylcellulose, polysorbate 80, hydroxyethylcellulose, gum arabic, tragacanth gum powder, sodium carboxymethylcellulose, polyoxyethylene sorbitan monolaurate, poloxamer, hydroxypropyl methylcellulose (HPMC), and sodium alginate.

[0039] Examples of the solubilizing agent include polyoxyethylene hydrogenated castor oil, polysorbate 80, niacinamide, polyoxyethylene sorbitan monolaurate, macrogol, glycerol fatty acid ester, lipoamino acid, and polyethylene glycol.

[0040] Examples of stabilizers include sodium sulfite and sodium metasulfite, examples of isotonicity adjusters include glycerin and sodium chloride, and examples of preservatives include methylparaben, ethylparaben, and sorbic acid.

[0041] The pH when the lyophilized preparation is dissolved in water is preferably 3 to 6, more preferably 3 to 5, further preferably 3 to 4.5, and particularly preferably 3.5 to 4.5.

[0042] The lyophilized preparation can be produced by a conventionally known method for producing a lyophilized preparation, for example, a method in which the preparation is frozen at a temperature of -25°C or lower, and then dried while the temperature is raised to room temperature while maintaining the vacuum degree at about 20 Pa or lower.

[0043] The injection of the present invention may be a lyophilized preparation. Such a lyophilized preparation may be dissolved in one or two or more solvents such as distilled water for injection, infusion solution, electrolyte solution, etc., to be used as an injection that dissolves when used.

[0044] The pharmaceutical composition of the present invention can be used for the treatment of specific diseases. Examples of the target diseases include fibroma, lipoma, myxoma, chondroma, osteoma, hemangioma, hemangioendothelioma, lymphoma, myeloma, myelosarcoma, reticular cell tumor, reticular cell sarcoma, melanoma, myoma, neuroma, glioma, schwannoma, sarcoma, osteosarcoma, myosarcoma, fibrosarcoma, papilloma, adenoma, cyst, brain tumor, cervical cancer, tongue cancer, pharyngeal cancer, Laryngeal cancer, thyroid cancer, esophageal cancer, lung cancer, breast cancer, gastric cancer, small intestine cancer such as duodenum, jejunum, ileum, colon cancer, cecum cancer, rectal cancer, large intestine cancer such as bladder cancer, kidney cancer, gallbladder cancer, prostate cancer, uterine cancer, cervical cancer, ovarian cancer, mesothelioma, head and neck cancer, adrenal cancer, gastrointestinal stromal tumors, appendix cancer, bile duct cancer, pancreatic cancer, liver cancer, skin cancer, choriocarcinoma, metastatic cancer, invasive cancer and other cancers, as well as their mixed tumors, metastatic tumors, etc.

[0045] Among them, bile duct cancer, colorectal cancer, pancreatic cancer, esophageal cancer, breast cancer, lung cancer, prostate cancer, bladder cancer, brain tumors, gastric cancer, liver cancer, skin cancer, chorionic carcinoma, kidney cancer, head and neck cancer, tongue cancer, metastatic cancer and invasive cancer are preferred, and bile duct cancer, colorectal cancer, pancreatic cancer, esophageal cancer and breast cancer are further recommended.

[0046] Dosage regimen The dosage regimen for the pharmaceutical composition of the present invention will depend on a variety of factors, including the type, race, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; and the patient's liver and kidney function. A doctor can easily determine the effective amount of the drug required to prevent, stop or stop the progression of symptoms and prescribe it.

[0047] In the pharmaceutical composition of the present invention, the dosage of the active ingredient can be appropriately selected according to the degree of symptoms, the age, sex, weight, sensitivity difference of the patient, the administration period, the administration interval, etc. From the viewpoint of effectiveness and safety, a single dose of 1 mg / m 2 ~60 mg / m 2 (body surface area), preferably 12.5 mg / m 2 ~60 mg / m 2 , 12.5mg / m 2 ~25 mg / m 2 , or 10 mg / m 2 ~40 mg / m 2 (body surface area), 25 mg / m is particularly recommended 2 In addition, depending on the symptoms, the dose can be reduced to 12.5 mg / m 2 wait.

[0048] Examples of dosage regimens for the pharmaceutical composition of the present invention include: - administering the pharmaceutical composition in a cycle of continuous administration for a constant time followed by a constant withdrawal time, - administering the pharmaceutical composition in a cycle of 5 consecutive days followed by 9 days of rest for a total of 14 days, - continuous intravenous administration of a constant amount of a pharmaceutical composition for a constant period of time, - 100 mL of the pharmaceutical composition is administered continuously intravenously over 90 minutes, - 1 to 60 mg / m 2 administering medication to a patient, - 12.5 to 60 mg / m 2 administering medication to a patient, - 12.5 to 25 mg / m 2 administering medication to a patient, -25 mg / m 2 administering medication to a patient, Etc., they can be combined and applied as needed.

[0049] The pharmaceutical composition of the present invention can exert excellent effects when administered to patients with cancers and the like having a non-rapid (slow and / or intermediate) type NAT2 gene (i.e., slow acetylation type and intermediate type caused by NAT2). In the present specification, "non-rapid NAT2 gene" refers to the intermediate and slow phenotypes of the three phenotypes of the acetylation rate of the NAT2 gene, namely, the rapid, intermediate and slow types.

[0050] For the 7 SNPs representing the polymorphism of the NAT2 gene (481C>T, 282C>T, 857G>A, 803A>G, 341T>C, 590G>A, 191G>A): the acetylation phenotype of NAT2 was analyzed using the combination of 2-SNP, 3-SNP and 4-SNP (Table 2-1), the SNP-free (standard) haplotype consisting of NAT2*4 homozygotes was considered to be a rapid acetylation type (Rapid), the heterozygotes of any haplotype mutation were considered to be a type with an intermediate acetylation speed (Intermediate), and the homozygotes of all haplotype mutations or the combination of heterozygotes of two or more haplotype mutations were considered to be a slow acetylation type (Slow) (Pharmacogenomics, Vol. 13: pp. 31-41, 2012). Here, the "NAT2*4" allele refers to the haplotype without SNP. In addition, according to Pharmacogenomics, Vol. 13: pp. 31-41, 2012, regarding the rapid type, in addition to "NAT2*4", patients with "NAT2*11, NAT2*12, NAT2*13" that do not cause amino acid mutations are also classified as rapid type.

[0051] Specifically, the phenotype of NAT2 acetylator inferred by two SNPs was determined by the analysis of two SNPs: rs1041983 (282C>T) and rs1801280 (341T>C). The phenotype of NAT2 acetylator inferred by three SNPs was determined by the analysis of three SNPs: rs1799929 (481C>T), rs1799930 (590G>A), and rs1799931 (857G>A). The phenotype of NAT2 acetylator inferred by four SNPs was determined by the analysis of four SNPs: rs1801279 (191G>A), rs1801280 (341T>C), rs1799930 (590G>A), and rs1799931 (857G>A).

[0052] If JPH203 is acetylated by NAT2, its inhibitory activity against cancer cells decreases. However, if the patient mainly has a NAT2 gene phenotype that is difficult to be acetylated, that is, if the NAT2 gene of a cancer patient has an intermediate or slow acetylation rate phenotype (Non-Rapid NAT2 gene), the acetylation rate of JPH203 becomes slow, so it is difficult to be acetylated, and the activity of JPH203 can be sustained for a long time.

[0053] The analysis of NAT2 gene polymorphism in the present invention can be performed by conventionally known methods, for example, by extracting DNA from the blood of a subject, treating the DNA using a microarray method, and then reading the genotype for examination, followed by data analysis.

[0054] Therefore, for the purpose of predicting the safety and efficacy of cancer therapeutic drugs and providing the therapeutic drugs to appropriate cancer patients, the administration of the pharmaceutical composition of the present invention to cancer patients is preferably carried out as follows.

[0055] Blood was collected from human patients with or suspected of having cancer, DNA was extracted from the blood samples, and the NAT2 gene polymorphism was confirmed. For patients whose NAT2 genetic polymorphism was judged to be non-rapid type (i.e., judged to have an acetylation rate of intermediate or slow), 25 mg / m 2 100 mL of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine sulfobutyl ether-cyclodextrin complex was continuously intravenously administered at a concentration of 1.5 mmol / l. At this time, the pharmaceutical composition was administered for 14 days in total, including continuous administration for 5 days and then 9 days of drug withdrawal as one cycle.

[0056] Furthermore, in the NAT2 gene diagnosis of a patient, by determining whether or not the patient has a non-rapid NAT2 gene, it is possible to predict whether O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine will have an effect on a patient with a specific disease, such as cancer.

[0057] Furthermore, it is possible that the therapeutic effect of cancer or the like may be improved in animal models or animal species in which the metabolic capacity of NAT2 is suppressed compared to animal models in which the metabolic capacity of NAT2 is normal. This may also be applicable to humans.

[0058] Furthermore, by applying the administration method related to the pharmaceutical composition of the present invention to patients, it is possible to provide excellent treatment for specific diseases, particularly cancer, by improving the safety of the drug and the effectiveness of the effect.

[0059] Furthermore, as a further embodiment of the present invention, a pharmaceutical composition for use in combination with NAT2 gene diagnosis is provided, which comprises O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof. The pharmaceutical composition can improve the effect when used for the treatment of specific diseases, particularly cancer, etc.

[0060] It should be noted that, as described above, NAT2 gene diagnosis is to perform an examination of NAT2 gene polymorphism to determine whether or not a non-rapid NAT2 gene is present. NAT2 gene diagnosis can be performed by a conventionally known diagnostic method, such as an analytical method for NAT2 gene polymorphism described later. Alternatively, NAT2 gene diagnosis can utilize the services of an inspection agency such as a company that provides gene inspection and gene diagnosis.

[0061] Alternatively, NAT2 gene diagnosis can also be performed using a NAT2 gene diagnosis kit. The NAT2 gene diagnosis kit is a collection of sample collection tools and reagents required for implementing the NAT2 gene polymorphism analysis method described below. Examples of reagents included in the kit include PCR enzyme solution, primers / probes, dH 2 O, positive control DNA, and dedicated buffer for control dilution, etc.

[0062] As another embodiment, the pharmaceutical composition of the present invention comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof can be used in combination with NAT2 gene diagnosis or a gene diagnosis kit. By using them in combination, the effect of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine on specific diseases, especially the effect of treating cancer, can be improved.

[0063] As a further embodiment, a method for effectively treating specific diseases, such as cancer, is provided by combining a pharmaceutical composition comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof of the present invention with a NAT2 gene diagnosis or a gene diagnosis kit.

[0064] For example, the pharmaceutical composition of the present invention comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof can be combined with the determination of NAT2 gene polymorphism and administered to a subject suffering from cancer according to the following dosage regimen.

[0065] (1) identifying and selecting the subject having a non-fast (slow and / or intermediate) type of NAT2 gene; (2) administering the pharmaceutical composition to the subject identified as having a non-fast (slow and / or intermediate) type of NAT2 gene.

[0066] In addition, the pharmaceutical composition comprising O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof of the present invention can be used in combination with a NAT2 inhibitor to treat a disease such as cancer. As a NAT2 inhibitor, for example, acetaminophen can be exemplified.

[0067] JPH203 is rapidly N-acetylated by NAT2 in the hepatocyte matrix to become Nac-JPH203. However, it is known that Nac-JPH203 has lower selectivity and activity for LAT1 than JPH203. By using it in combination with a NAT2 inhibitor, the acetylation of JPH203 is inhibited, thereby maintaining the effect of JPH203.

[0068] The dosage of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof and the NAT2 inhibitor can be respectively used in effective amounts, and can be appropriately selected according to the subject, administration route, disease, combination, etc.

[0069] The administration time of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof and the NAT2 inhibitor is not particularly limited. O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof and the NAT2 inhibitor of the present invention may be administered to the subject simultaneously or at a time difference.

[0070] By using O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof in combination with a NAT2 inhibitor, it is possible to reduce the dosage of each, extend the treatment time, and obtain a synergistic effect.

[0071] Combination with Boron Neutron Capture Therapy (BNCT) Furthermore, the O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof of the present invention can be used in combination with boron neutron capture therapy for treating cancer.

[0072] Examples of compounds used for boron neutron capture therapy include boronated amino acids, and boronated phenylalanine (BPA) (trade name: Bolofilan B10) is particularly recommended. Boron neutron capture therapy is the combination of boronated amino acids and neutron beam irradiation. In this method, boronated amino acids (such as boronated phenylalanine BPA) are injected into the body and taken up by cancer cells. At this time, BPA is taken up by cancer cells through LAT1. In contrast, if neutron rays are irradiated, nuclear division occurs in cancer cells, causing cell destruction. However, since BPA is easily excreted from cancer cells through LAT1, a large amount of BPA must be administered. On the other hand, if BPA is administered in large quantities, normal cells will also take up BPA. As a result, it affects the surrounding normal cells, causing side effects, and also affects the kidneys, an organ for metabolism and excretion.

[0073] Therefore, by using the O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine of the present invention in combination with boron neutron capture therapy, BPA can be effectively enclosed in cancer cells, thereby reducing the dosage of BPA and thus reducing the effect (= side effect) on normal cells (Jutabha et al. Journal of Pharmacological Sciences, Volume 130, Issue 3, Supplement (Proceedings of the 89th Annual Meeting of the Japanese Pharmacological Society) 3-0-84).

[0074] That is, as described in the literature of Jutabha et al., BPA is taken into cells through LAT1 on the surface of cancer cells. In this state, BPA is also excreted from the cells by LAT1. By administering JPH203 after BPA is absorbed by cancer cells and before it is excreted from cancer cells, LAT1 can be blocked and BPA can be sealed into cancer cells (the time difference between the administration of BPA and JPH203 becomes important). Therefore, even if the dosage of BPA is small, cancer cells can be attacked efficiently. In other words, JPH203 is effective in blocking BPA. Nac-JPH203 has a lower cell proliferation inhibitory effect than JPH203, so it is believed that the effect of NAT2 non-rapid type is higher than that of NAT2 rapid type.

[0075] The current standard method of BPA administration is intravenous drip administration, which is administered at a rate of 200 mg / kg / h for 2 hours before neutron irradiation. During irradiation, 100 mg / kg is administered at a constant rate of about 100 mg / kg / h in accordance with the end time of irradiation. In order to maintain a high concentration of BPA in cancer cells, the target is over-set, so the concentration of BPA in the extracellular matrix becomes higher, and the impact on normal cells also increases. The timing of JPH203 administration is as follows: BPA is administered by intravenous drip at a rate of 100-200 mg / kg / h for 30 minutes to 1 hour before neutron irradiation, and then immediately at 10-25 mg / m 2JPH203 was administered by intravenous drip at a rate of / h. Neutron beam was irradiated 15 to 30 minutes after the start of administration of JPH203.

[0076] It should be noted that if the local concentration of BPA becomes high, it is also taken up into normal cells through LAT2 on the surface of normal cells, but the uptake efficiency of LAT2 is lower than that of LAT1. Since JPH203 only blocks LAT1, BPA in cancer cells is continuously blocked at a sufficiently high concentration at the beginning of neutron irradiation.

[0077] When O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutical composition comprising the compound of the present invention is used in combination with boron neutron capture therapy, the following dosage regimen is recommended.

[0078] For example, BPA is intravenously infused into cancer patients at a rate of 200 mg / kg per hour for 1 hour. The administration of BPA is then stopped and immediately replaced with 10-20 mg / m 2 / h began intravenous administration of JPH203, neutron beam irradiation began 30 minutes later, and intravenous drip administration of JPH203 continued until the end of irradiation.

[0079] Furthermore, the combination of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine and boron neutron capture therapy of the present invention may be combined with the method for measuring the NAT2 gene polymorphism.

[0080] As such a method, the following dosage regimens can be mentioned: (1) identifying and selecting the subject having a non-fast (slow and / or medium) type NAT2 gene; (2) administering a boron-containing compound to the subject identified as having a non-rapid (slow and / or intermediate) type of NAT2 gene before administering the pharmaceutical composition; (3) administering the pharmaceutical composition to the subject.

[0081] Example Although the present invention is described below through specific embodiments, it should be understood that the present invention is not limited to these embodiments and that various changes and modifications may be made thereto by those skilled in the art without departing from the scope or spirit of the present invention defined in the appended claims.

[0082] Example 1 Drug used: Dissolve JPH203-SBECD (active ingredient 50 mg) in 9.7 ml of water for injection to a concentration of 50 mg / 10 ml, and finally make a total volume of 100 ml according to the patient's body surface area.

[0083] Patients with advanced solid cancer who were refractory or intolerant to standard treatment were enrolled to evaluate the efficacy of JPH203-SBECD (12, 25, 40, 60, and 85 mg / m 2 In order to determine the safety of ) and determine the recommended dose for the next phase and analyze its pharmacokinetics, a non-controlled, open-label Phase I trial was conducted in Japan. The study drug was set at 12 mg / m 2 The initial dose was set according to the Fibonacci transformation and 5 levels (12, 25, 40, 60, 85 mg / m 2 ). Three subjects who registered within 28 days after obtaining consent were selected as subjects for each level, and a single dose was administered within 4 days after registration. After the first subject received a single dose, the second and subsequent subjects were started at a time when safety was confirmed to be ok based on the results of the pre-dose check on the first day of the first 7-day repeated dosing (cycle 1).

[0084] The subjects who received a single dose and were judged to have no problems with their health were selected as subjects, and the cycle of repeated administration of the study drug continued from 7 days after the single dose until the discontinuation criteria were reached. In addition, the administration of the first subject was started at the time when safety was confirmed to be ok 49.5 hours after the start of administration on the 7th day of cycle 1, and the administration of the second and subsequent subjects was started.

[0085] In single administration and cycle 1 (once a day, 7 days), the subjects were continuously intravenously administered the study drug for 90 minutes through a drip circuit containing a final filter (pore size 0.22μm) and a metering pump. The study drug was prepared in a way that the total amount was 100mL of an appropriate infusion such as physiological saline, Ringer's solution, etc., and the target dosage was achieved. In cycle 2 and beyond, the subjects were similarly administered the study drug once a day for 7 days, with the same dosage and dosage as cycle 1. The study drug was prepared in an appropriate infusion such as physiological saline, Ringer's solution, etc., so as to achieve the target concentration.

[0086] As a result, among the 5 patients with biliary tract cancer, 1 case of partial response (PR) and 2 cases of stable disease (SD) were confirmed (total response rate 20.0%, disease control rate 60.0%), suggesting the effectiveness of this drug for patients with biliary tract cancer. In terms of safety, no clinically problematic adverse events or side effects were observed. Based on these results, it is believed that there is no problem with tolerability, and it can be expected to have a constant effectiveness for patients with biliary tract cancer.

[0087] The results of the pharmacokinetic parameter analysis obtained in the Japanese Phase I trial confirmed that the individual differences in the concentration of N-acetylated form (Nac-JPH203) in blood and urine were large among subjects who were given the same dose per unit body surface area. After intravenous administration, JPH203 is taken up into hepatocytes via organic anion transporters (OATPs), which are liver transporters. It is believed that JPH203 taken up by hepatocytes is converted into Nac-JPH203 by NAT2 in hepatocytes and excreted into bile via biliary excretion transporters. Therefore, it is speculated that the individual differences in the concentration of Nac-JPH203 in urine are caused by differences in the rate of acetylation of NAT2, which is believed to be responsible for most of the in vivo metabolism of JPH203.

[0088] Pharmacokinetic analysis Blood (whole blood) and urine were collected from the subjects. Whole blood used for concentration measurement was collected 0.5, 1, 1.5, 2, 3, 4.5, 6, 12, 25.5, and 49.5 hours after the start of administration. Urine used for concentration measurement was collected by urine accumulation from 0-3, 3-6, 6-12, and 12-25.5 hours after the start of administration. The concentration of JHP203 in blood and urine and the concentration of Nac-JPH203 as its acetyl form were measured using a verified LC-MS / MS measurement system.

[0089] The pharmacokinetic parameters were calculated using the blood JPH203 concentration and the Nac-JPH203 concentration. Among them, the ratio of Nac-JPH203 to JPH203 in the blood of each subject was calculated using AUC0-∞. In addition, for each subject, the sum of the masses of JPH203 and Nac-JPH203 contained in the urine collected from the start of administration to 25.5 hours was calculated, and the ratio of Nac-JPH203 to JPH203 in the urine of each subject was calculated.

[0090] Determination of NAT2 gene polymorphism The measurement of NAT2 gene polymorphism can be performed by the following real-time fluorescence quantitative PCR (Real-Time PCR) method or the method using Luminex System. In either method, the following 7 types of SNPs are analyzed for NAT2.

[0091]

[0092] Real-time fluorescence quantitative PCR The measurement was performed by real-time fluorescence quantitative PCR according to the following method. It should be noted that the details of the protocol can be changed as appropriate.

[0093] (i) After measuring the DNA concentration of NAT2, two upstream primers and one downstream primer were designed that could amplify a region containing seven types of SNPs to be analyzed.

[0094] (ii) These PCR primers are used to study PCR conditions and determine the final PCR primer set and PCR conditions to be used. Then, the genomic DNA is subjected to sequence analysis based on the Sanger sequencing method according to the determined conditions.

[0095] (iii) After PCR, the PCR product is purified and sequenced using a capillary sequencer. Sequencing primers are used to sequence the PCR product from both sides.

[0096] How to use the Luminex system The measurement was performed using the Luminex system method according to the following method. It should be noted that the details of the protocol may be changed as appropriate.

[0097] (i) Probe solid phase beads corresponding to the seven SNPs to be analyzed by NAT2 were prepared.

[0098] (ii) The NAT2 gene is amplified using a biotin-labeled primer. The PCR amplification product is bound to a probe and fluorescently labeled with SA-PE.

[0099] (iii) Data were acquired using the Luminex system and the NAT2 gene type was displayed in the analysis software.

[0100] Analysis of NAT2 gene polymorphism The examination of NAT2 gene polymorphism was carried out as follows. DNA was extracted from blood samples during the Phase I trial, and the phenotype was classified based on the information of each SNP according to the NAT2 gene polymorphism. The outline of the method is as follows: DNA extraction was performed using QIAamp DNA BloodMini kit (registered trademark) (QIAGEN Co., Ltd.) after 200 μL of whole blood was dissolved in 100 μL of AE buffer. The array and haplotype associated with the SNP at the site encoding NAT2 were measured using an ultra-micro spectrophotometer (Nano Drop 2000 (registered trademark), Thermo Fisher Scientific) (the concentration was adjusted to 5 ng / μL). TaqMan (registered trademark) SNP Genotyping assay (reaction volume 5 μL) was used for gene polymorphism analysis. PCR primers and fluorescent probes required for SNP detection were designed using Primer Express (trademark) (Applied Biosystems, CA, USA). The fluorescent probe was labeled with a reporter dye (carboxyfluorescein or VIC (registered trademark), Applied Biosystems) and reacted specifically with one of the two bases in seven NAT2 SNPs (rs1801279 (191G>A), rs1041983 (282C>T), rs1801280 (341T>C), rs1799929 (481C>T), rs1799930 (590G>A), rs1208 (803A>G), and rs1799931 (857G>A)). The PCR reaction conditions were 95°C for 10 minutes, followed by 92°C for 15 seconds and 60°C for 90 seconds for 50 cycles.

[0101] The gene of 16 patients who participated in the phase I trial was analyzed for 7 SNPs representing NAT2 gene polymorphisms (481C>T, 282C>T, 857G>A, 803A>G, 341T>C, 590G>A, and 191G>A).

[0102] The acetylation phenotype of NAT2 was analyzed using a combination of 2-SNP, 3-SNP, and 4-SNP (Table 2-1). The results showed that the rapid acetylation type (Rapid) was composed of homozygotes of the standard haplotype NAT2*4, the intermediate acetylation type (Intermediate) was composed of heterozygotes of any haplotype mutation, and the slow acetylation type (Slow) was composed of homozygotes of the mutant haplotype or a combination of heterozygotes of two or more mutant haplotypes (Table 2-2).

[0103]

[0104]

[0105] Classification of phenotypes based on NAT2 genotyping The SNP phenotypes of NAT2 in the 16 subjects were consistent regardless of the type of cancer (colorectal cancer, biliary tract cancer, pancreatic cancer, esophageal cancer, and breast cancer), and any combination of 2-SNP, 3-SNP, and 4-SNP was consistent. The details of the results are shown in Table 3.

[0106]

[0107] SD: Stable (a state in which the tumor size does not change) PD: Partially effective (the sum of tumor sizes increases by more than 20% and the absolute value increases by more than 5 mm, or new lesions appear) PAAD: Pancreas Adenocarcinoma CHOL: Cholangiocarcinoma COAD: Colon Adenocarcinoma BRCA: Breast Adenocarcinoma ESCA: Esophageal Carcinoma Relationship between NAT2 phenotype and clinical outcomes (safety, efficacy) If the deviation of the Nac-JPH203 / JPH203 concentration ratio of the 16 individuals is classified into the fast type, the intermediate type, and the slow type of NAT2, the type with more metabolites Nac-JPH203 is the fast type of NAT2, and the individuals with less metabolites are the slow type of NAT2. In these NAT2 fast groups, an increase in liver function test values ​​was confirmed in 4 cases with a high Nac-JPH203 ratio, and 2 of them were judged to be DLT (Dose Limiting Toxicity). These patients were 2 cases of colorectal cancer, 1 case of esophageal cancer, and 1 case of pancreatic cancer, and no patients with biliary tract cancer were included.

[0108] That is, in cancer patients with NAT2 fast type, the value of liver dysfunction caused by JPH203 administration was high.

[0109] The clinical efficacy of JPH203 was evaluated by DCR (Disease Control Rate), and the results showed that among all 16 cases, the number of patients who improved (PR or SD) was 1 case (12.5%) in the NAT2 fast group of 8 cases and 4 cases (50%) in the NAT2 non-fast (slow and moderate) group. Among patients with biliary tract cancer (BDC: 5 cases in total), 0 cases (0%) in the NAT2 fast group and 3 cases (75%) in the non-fast group showed improvement (Table 4).

[0110] Regarding progression-free survival (PFS), the survival curve was prepared using the Kaplan-Meier method, and the median of progression-free survival and its 95% confidence interval were calculated. The median result was 37.0 days (CSR 11.4.1 effectiveness analysis). Among them, the median PFS of the NAT2 rapid group was 32 days, and that of the NAT2 non-rapid group was 58 days. Moreover, the median PFS of 4 of the 5 patients with biliary tract cancer in the NAT2 non-rapid group was 99.5 days. Regarding overall survival (OS), the median of the 16 cases was 3.5 months, of which the NAT2 rapid group was 2.5 months, and the NAT2 non-rapid group (4 cases of biliary tract cancer) was 6 months. There was a significant difference between the two groups.

[0111]

[0112] JPH203 is taken up by the liver and metabolized. JPH203 taken up by the liver is acetylated and converted to N-acetyl-JPH203 (Nac-JPH203), and Nac-JPH203 is excreted from the body depending on the blood flow rate. Both JPH203 and Nac-JPH203 are taken up by hepatocytes via OATP (organic anion transporter), and excreted from hepatocytes via Mrp2 (multidrug resistance associated protein 2). The uptake rate of JPH203 (Nac-JPH203) by OATP1B1 is lower than the excretion rate of JPH203 (Nac-JPH203) by Mrp2, so it is believed that the uptake process (rats) is the rate-limiting step for JPH203 metabolism. However, in humans, due to the relationship with the blood flow rate of the liver, much JPH203 is retained in hepatocytes, and it is believed that the metabolism in cells after uptake (the process of acetylation by NAT2) is the rate-limiting step.

[0113] Analysis of NAT2 in subjects participating in the Phase I trial using 7 SNPs revealed that the NAT2 phenotype (acetylation rate: fast, medium, and slow) of each subject was consistent for any combination of 2-SNP, 3-SNP, and 4-SNP (Table 3).

[0114] The Nac-JPH203 / JPH203 ratios of the 17 patients who participated in the Phase I trial were roughly correlated in the blood and urine, so it is presumed that JPH203 is metabolized in the liver and excreted into the blood in accordance with its concentration, and passively excreted into the urine from the kidneys (it should be noted that one case was excluded because the concentration results in urine could not be used at the stage of analysis). All 16 cases with high Nac-JPH203 / JPH203 ratios were patients with NAT2 rapid type. Among them, in the top 4 cases with high Nac-JPH203 production ratios, an increase in liver function test values ​​was confirmed, and 2 of them were judged as DLT. There are many unclear parts about the toxicity of Nac-JPH203 metabolites, but in a study in which JPH203 was repeatedly administered to NAT2-deficient dogs for 4 weeks, compared with rats and humans, the histological changes in the liver, kidneys, etc. were milder than those in rats, which is a result that supports the clinical results.

[0115] On the other hand, regarding effectiveness, one of the eight cases in the NAT2 rapid group (colorectal cancer) showed SD (12.5%), while four of the eight cases in the NAT2 non-rapid group showed PR or SD (50%). The overall survival of the NAT2 non-rapid group was significantly longer than that of the NAT2 rapid group, and the median progression-free survival (PFS) was about twice as long in the NAT2 rapid group as in the NAT2 non-rapid group. Among the four cases that showed effectiveness, three were biliary tract cancers, indicating that the metabolism of NAT2 progresses slowly, and the effectiveness of JPH203 may be more effectively expressed in tissues where the local concentration of JPH203 remains constant or above. In the NAT2 rapid group, cases 102 and 403 eventually developed PD, but PFS was longer in the NAT2 rapid group. The Nac-JPH203 / JPH203 ratio in the blood of these cases was relatively low, and it is speculated that the safety and effectiveness of JPH203 are superior in the NAT2 non-rapid group.

[0116] Therefore, it is suggested that SNP analysis of NAT2 is useful as a "companion diagnostic" for identifying subjects (cancer patients, etc.) who are effective for treatment with JPH203 and maximizing its anti-tumor activity.

[0117] In addition, the safety and efficacy of JPH203 and the superiority of administration to patients with the non-rapid type NAT2 gene can be further confirmed by a randomized comparative controlled trial in which the treatment target patients are pre-classified into a group with the non-rapid type gene and a group with the rapid type gene and then tested.

[0118] In such a test, JPH203, for example, was administered at 25 mg / m 2The basic cycle is 14 days, with continuous administration for 5 days and rest for 9 days. The study drug allocated to each subject is administered to a total of 100 mL using a syringe pump or infusion pump through continuous intravenous administration over 90 minutes.

[0119] As a data analysis method, for example, for progression-free survival based on independent blind central review, all subjects who were given one dose of the study drug and had efficacy data were taken as the subjects, and the hazard ratio and its 95% confidence interval in each group (rapid type, non-rapid type) were calculated using the Cox proportional hazards model according to the metabolic type of NAT2 (rapid type, non-rapid type). Furthermore, the p-value relative to the interaction term can be calculated using the Cox proportional hazards model that includes the interaction term between the dosing group and the NAT2 metabolic type.

[0120] In addition, for overall survival, the hazard ratio and its 95% confidence interval in each group (rapid type, non-rapid type) were calculated using the Cox proportional hazards model according to the metabolic type of NAT2 (rapid type, non-rapid type), and the p-value for the interaction term was calculated using the Cox proportional hazards model including the interaction term between the dosing group and the metabolic type of NAT2. In addition, for progression-free survival based on physician evaluation, the hazard ratio and its 95% confidence interval for each group (rapid type, non-rapid type) were calculated using the Cox proportional hazards model according to the metabolic type of NAT2 (rapid type, non-rapid type), and the p-value for the interaction term was calculated using the Cox proportional hazards model including the interaction term between the dosing group and the metabolic type of NAT2.

[0121] Example 2 Combination trial of JPH203 and NAT2 inhibitors To evaluate whether administration of the NAT2 inhibitor acetaminophen (APAP) increases plasma and liver concentrations of JPH203 or decreases concentrations of its metabolite Nac-JPH203.

[0122] The animals used were 8-week-old male SD rats, which were administered with 1.20 nmol / (min•kg) of JPH203. The rats were diluted with physiological saline to this concentration, and the administration rate was 20 μL / min. 500 and 1500 mg / kg of acetaminophen were administered 30 minutes before the administration of JPH203. (0.5% methylcellulose aqueous solution). The blood collection time points were set at 0 (blank), 30, 60, 120, 180, and 240 minutes after the administration of JPH203. In addition, the sample volume collected was 0.2 mL of plasma, about 0.4 mL of blood, and liver, kidney, brain, and cerebrospinal fluid (CSF).

[0123] The results are shown in Figure 1 It was found that the combined use of JPH203 and NAT2 inhibitor significantly reduced the ratio of Nac-JPH203 / JPH203 in the blood of rats compared with the use of JPH203 alone, slowed down the metabolism of JPH203, and increased the blood concentration of JPH203, thus maintaining the effect of JPH203.

[0124] Example 3 Pancreatic cancer-derived T3M4 cells were used as representative cancer cells. BPA was added to the cells, and 10 μM JPH203 was added 15 minutes later to measure the concentration of BPA enclosed in the cells. The results are shown in Figure 2 In the case where JPH203 (●) was not added, BPA was excreted from the cells over time, so the intracellular concentration decreased. On the other hand, in the case where JPH203 (■) was added, the intracellular concentration was maintained because JPH203 inhibited the excretion of BPA. After 30 minutes, the intracellular concentrations of the two differed by about 10 times.

[0125] Industrial applicability The pharmaceutical composition of the present invention can provide treatment for cancer and the like with improved safety of the drug and improved effectiveness of the effect.

Claims

1. Use of a pharmaceutical composition in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: The pharmaceutical composition comprises O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof.

2. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: The cancer disease is selected from the group consisting of biliary tract cancer, colorectal cancer, pancreatic cancer, esophageal cancer, breast cancer, lung cancer, prostate cancer, bladder cancer, brain tumor, stomach cancer, liver cancer, skin cancer, chorionic carcinoma, kidney cancer, head and neck cancer, tongue cancer, metastatic cancer and invasive cancer.

3. Use of the pharmaceutical composition according to claim 2 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: The cancer disease is selected from the group consisting of biliary tract cancer, colorectal cancer, pancreatic cancer, esophageal cancer, and breast cancer.

4. Use of the pharmaceutical composition according to claim 3 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: The disease is bile duct cancer.

5. Use of the pharmaceutical composition according to any one of claims 1 to 4 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: The pharmaceutical composition is administered in a cycle of continuous administration for a constant time followed by a constant rest time.

6. Use of the pharmaceutical composition according to claim 5 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: The pharmaceutical composition was administered in a cycle of 5 consecutive days of administration followed by 9 days of rest for a total of 14 days.

7. Use of the pharmaceutical composition according to any one of claims 1 to 4 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: 1–60 mg / m 2 The pharmaceutical composition is administered to the subject once daily.

8. Use of the pharmaceutical composition according to claim 7 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: 12.5–60 mg / m 2 The pharmaceutical composition is administered to the subject once daily.

9. Use of the pharmaceutical composition according to claim 8 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: 12.5 to 25 mg / m 2 The pharmaceutical composition is administered to the subject once daily.

10. Use of the pharmaceutical composition according to any one of claims 1 to 4 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: A constant amount of the pharmaceutical composition is administered intravenously continuously over a constant period of time.

11. Use of the pharmaceutical composition according to claim 10 in the preparation of a drug for treating cancer diseases in patients with non-rapid NAT2 genes, characterized in that: 100 mL of the pharmaceutical composition was continuously intravenously administered over 90 minutes.

12. Use of a pharmaceutical composition in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: The pharmaceutical composition comprises O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof.

13. Use of the pharmaceutical composition according to claim 12 in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: The pharmaceutical composition is administered in a cycle of continuous administration for a constant time followed by a constant rest time.

14. Use of the pharmaceutical composition according to claim 13 in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: The pharmaceutical composition was administered in a cycle of 5 consecutive days of administration followed by 9 days of rest for a total of 14 days.

15. Use of the pharmaceutical composition according to claim 12 in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: 1–60 mg / m 2 The pharmaceutical composition is administered to the subject once daily.

16. Use of the pharmaceutical composition according to claim 15 in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: 12.5–60 mg / m 2 The pharmaceutical composition is administered to the subject once daily.

17. Use of the pharmaceutical composition according to claim 16 in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: 12.5 to 25 mg / m 2 The pharmaceutical composition is administered to the subject once daily.

18. Use of the pharmaceutical composition according to claim 12 in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 gene who are ineffective or intolerant to standard therapy, characterized in that: A constant amount of the pharmaceutical composition is administered intravenously continuously over a constant period of time.

19. Use of the pharmaceutical composition according to claim 18 in the preparation of a drug for treating advanced biliary tract cancer in which standard therapy is ineffective or intolerant to patients with non-rapid NAT2 gene, characterized in that: 100 mL of the pharmaceutical composition was continuously intravenously administered over 90 minutes.

20. Use of a pharmaceutical composition in the preparation of a drug for treating advanced biliary tract cancer in patients with non-rapid NAT2 genes who are ineffective or intolerant to standard therapy, characterized in that: The pharmaceutical composition comprises O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof and can reduce the occurrence of liver dysfunction.

21. Use of a pharmaceutical composition in preparing a drug for treating cancer, characterized in that: The active ingredients of the pharmaceutical composition consist of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof and a NAT2 inhibitor.

22. Use of the pharmaceutical composition according to claim 21 in preparing a drug for treating cancer diseases, characterized in that: The NAT2 inhibitor is acetaminophen.

23. Use of the pharmaceutical composition according to claim 20 or 21 in preparing a drug for treating cancer, characterized in that: The cancer disease is selected from the group consisting of biliary tract cancer, colorectal cancer, pancreatic cancer, esophageal cancer, breast cancer, lung cancer, prostate cancer, bladder cancer, brain tumor, stomach cancer, liver cancer, skin cancer, chorionic carcinoma, kidney cancer, head and neck cancer, tongue cancer, metastatic cancer and invasive cancer.

24. Use of the pharmaceutical composition according to claim 23 in preparing a drug for treating cancer diseases, characterized in that: The cancer disease is selected from the group consisting of biliary tract cancer, colorectal cancer, pancreatic cancer, esophageal cancer, and breast cancer.

25. Use of the pharmaceutical composition according to claim 24 in the preparation of a drug for treating cancer diseases, characterized in that: The disease is bile duct cancer.

26. Use of the pharmaceutical composition according to claim 25 in preparing a drug for treating cancer diseases, characterized in that: Biliary tract cancer is an advanced bile duct cancer that is refractory or intolerant to standard treatments.

27. Use of the pharmaceutical composition according to any one of claims 1, 20 and 21 in the preparation of a drug for treating cancer, characterized in that: After administration of the pharmaceutical composition, the cancer is further killed by neutron capture therapy.

28. A pharmaceutical composition, characterized in that The active ingredients of the pharmaceutical composition consist of O-(5-amino-2-phenylbenzoxazol-7-yl)methyl-3,5-dichloro-L-tyrosine or a pharmaceutically acceptable salt thereof and a NAT2 inhibitor.

29. The pharmaceutical composition according to claim 28, characterized in that The NAT2 inhibitor is acetaminophen.

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